LOCUS       sequence-7696-ex        5191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Deuschle K, Chaudhuri B, Okumoto S, Lager I, Lalonde S, Frommer WB
  TITLE     Rapid Metabolism of Glucose Detected with FRET Glucose Nanosensors 
            in Epidermal Cells and Intact Roots of Arabidopsis RNA-Silencing 
            Mutants.
  JOURNAL   Plant Cell. 2006 Sep . 18(9):2314-25.
  PUBMED    16935985
REFERENCE   2  (bases 1 to 5191)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2006 Sep . 18(9):2314-25."
COMMENT     Sequence Label: pRSET FLIPglu-600uDelta13 Aphrodite
FEATURES             Location/Qualifiers
     source          1..5191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             67..89
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..162
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(166..174,175..189)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(202..204,205..207,208..720)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(247..268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(508..527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     855..876
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2537..2555)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2551..2598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2695..3150
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2782..2801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2992..3013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3231..3335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3336..3404,3405..4196)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4367..4955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4856..4875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5109..5126
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gataattttg tttaacttta agaaggagat atacatatgc ggggttctca tcatcatcat
      121 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      181 gacgataagg atccgggccg catggtgagc aagggcgagg agctgttcac cggggtggtg
      241 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      301 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      361 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
      421 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      481 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      541 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      601 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
      661 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cgctgatact
      901 cgcattggtg taacaatcta taagtacgac gataacgcga tgtctgtagt gcgcaaggct
      961 attgagcaag atgcgaaagc cgcgccagat gttcagctgc tgatgaatga ttctcagaat
     1021 gaccagtcca agcagaacga tcagatcgac gtattgctgg cgaaaggggt gaaggcactg
     1081 gcaatcaacc tggttgaccc ggcagctgcg ggtacggtga ttgagaaagc gcgtgggcaa
     1141 aacgtgccgg tggttttctt caacaaagaa ccgtctcgta aggcgctgga tagctacgac
     1201 aaagcctact acgttggcac tgactccaaa gagtccggca ttattcaagg cgatttgatt
     1261 gctaaacact gggcggcgaa tcagggttgg gatctgaaca aagacggtca gattcagttc
     1321 gtactgctga aaggtgaacc gggccatccg gatgcagaag cacgtaccac ttacgtgatt
     1381 aaagaattga acgataaagg catcaaaact gaacagttac agttagatac cgcaatgtgg
     1441 gacaccgctc aggcgaaaga taagatggac gcctggctgt ctggcccgaa cgccaacaaa
     1501 atcgaagtgg ttatcgccaa caacgatgcg atggcaatgg gcgcggttga agcgctgaaa
     1561 gcacacaaca agtccagcat tccggtgttt ggcgtcgatg cgctgccaga agcgctggcg
     1621 ctggtgaaat ccggtgcact ggcgggcacc gtactgaacg atgctaacaa ccaggcgaaa
     1681 gcgacctttg atctggcgaa aaacctggcc gatggtaaag gtgcggctga tggcaccaac
     1741 tggaaaatcg acaacaaagt ggtccgcgta ccttatgttg gcgtagataa agacaacctg
     1801 gctctcgagt tgtttacggg cgtcgtcccg atcctcgtgg aactcgacgg ggatgttaac
     1861 gggcataagt tttcggtcag cggggaaggg gagggggacg cgacgtatgg gaagctcact
     1921 ctcaagctga tctgtacgac ggggaaactc ccggtcccgt ggccgacgct ggtcacgacg
     1981 ctgggatacg ggctccaatg ctttgcgagg tatccggacc acatgaaaca gcatgacttt
     2041 ttcaaatcgg cgatgccgga gggatacgtg caggaacgga cgatcttttt caaagacgat
     2101 gggaactata agacgcgggc ggaagtcaag tttgaagggg acacgctcgt caaccggatc
     2161 gaactcaagg ggattgactt caaagaggat gggaacatac tcggccataa gctcgaatac
     2221 aattacaact cgcataacgt atacatcacc gcggataagc aaaagaatgg gatcaaagcc
     2281 aatttcaaaa tccggcataa catagaggat gggggggtcc aactggcgga tcactatcag
     2341 caaaacacgc cgatagggga tgggccggtc ctcctcccgg ataaccatta cctctcgtac
     2401 caaagcgcgc tctcgaagga cccgaatgag aaacgggacc acatggttct cctggagttc
     2461 gtcacggcgg cgggcatata gaagcttgat ccggctgcta acaaagcccg aaaggaagct
     2521 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2581 gtcttgaggg gttttttgct gaaaggagga actatatccg gatctggcgt aatagcgaag
     2641 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     2701 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2761 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2821 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagagctt
     2881 tacggcacct cgaccgcaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2941 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3001 tgttccaaac tggaacaaca ctcaacccta tcgcggtcta ttcttttgat ttataaggga
     3061 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaata tttaacgcga
     3121 attttaacaa aatattaacg tttacaattt cgcctgatgc ggtattttct ccttacgcat
     3181 ctgtgcggta tttcacaccg catacaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3541 cttttaaagt tctgctatgt gatacactat tatcccgtat tgacgccggg caagagcaac
     3601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3841 aagccatacc aaacgacgag agtgacacca cgatgcctgt agcaatgcca acaacgttgc
     3901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgaa
     3961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4081 cagatggtaa gcgctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4501 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4981 ggttcctggg cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5161 tccccgcgcg ttggccgatt cattaatgca g
//
